Tissue regeneration of salivary gland by the growth factor and cultured cells.

1.本实验研究了碱性成纤维细胞生长因子在正常大鼠和人颌下腺细胞单层培养中的作用。以3T3细胞为饲养层，培养大鼠和福曼颌下腺上皮细胞。观察不同浓度的碱性成纤维细胞生长因子对培养细胞传代的影响。在大鼠和人的细胞中，BrdU阳性细胞的百分率逐渐增加到50 ng/ml，然后在100 ng/ml时急剧增加，但当浓度高于100 ng/ml时，BrdU阳性细胞的百分率达到最高。在大鼠和人细胞中，从培养第4天起，100 ng/mlbFGF组的细胞总数明显高于对照组，而培养的细胞形态与加入bFGF者无明显差异。这些结果表明，碱性成纤维细胞生长因子对唾液腺上皮细胞的主要作用是作为有丝分裂刺激。以萎缩的大鼠颌下腺为模型，观察碱性成纤维细胞生长因子对组织修复的促进作用。在手术显微镜下仔细分离腺管与相关血管和神经，并用8-0线结扎。结扎后2周，腺体组织严重萎缩，重量减轻。然后，取下结扎线，通过导管逐步注入不同剂量的碱性成纤维细胞生长因子、异丙肾上腺素或盐水。异丙肾上腺素和碱性成纤维细胞生长因子均能显著促进细胞增殖。免疫组织化学结果显示受损腺体中ANI-FGF1型受体抗体的免疫反应性增强，这可能与受损腺体与正常腺体对bFGF反应的不同有关。这些结果表明，碱性成纤维细胞生长因子能够促进唾液腺的组织修复。

1. In this study, the effects of bFGF were investigted in monclayer culture of normal rat and human submandibular gland cells. Epithelial cells from rat and fuman submandibular glands were cultivated wife the aid of 3T3 cells as a feeder layer. The effects of different concentrations of bFGF on the second passage of these cultured cells were examined. In both the rat and human cells, the percentage of bromodeoxyuridine (BrdU)-positive cells gradually increased up to 50 ng/ml, and then increased sharply at 100ng/ml. However, at concentrations higher than 100ng/ml, the peroentage of BrdU-positive cells reached a pleteau. In both rat and human cells, total cell nembers at 100ng/ml bFGF were sihnificantly higher than those of the oontrol group from culture day 4. On the other hand, the morphology of the cultured cells showed no difference either with or without bFGF. These results indicate that a major effect of bFGF on salivary gland eppithelial cells is to act as a mitogenic stimulus.2. A model of atrophic rat submandibular gland was used to examine the ability of bFGF to accelerated tissue repair. the gland duct was separated carefully from associated blood vessels and nerve, and ligated with a 8-0 suture under a surgical microscope. Two weeks after ligation,the glandular tissue showed severe atrophy and weight loss. Thereafter, the ligature was removed and various amounts of bFGF, isoproterenol or saline were instilled letrogradely through the duct. Both isoproterenol and bFGF increased cell proliferation significantly. The results from immunohistchemcal tests against ani-FGF receptor-type 1 antibody demonstrated increased immunoreactivity in the damaged gland, which might be involved in the difference in the response to bFGF between damaged and normal glands. These results indicated that bFGF can accelerate tissue repair in salivary gland.